2015 White Paper on Recent Issues in Bioanalysis: Focus on New Technologies and Biomarkers (Part 2 – Hybrid LBA/LCMS and Input from Regulatory Agencies)

Abstract

The 2015 9th Workshop on Recent Issues in Bioanalysis (9th WRIB) took place in Miami, Florida with participation of over 600 professionals from pharmaceutical and biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. It is once again a 5-day week long event - a full immersion bioanalytical week - specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest in bioanalysis. The topics covered included both small and large molecules, and involved LCMS, hybrid LBA/LCMS, LBA approaches including the focus on biomarkers and immunogenicity. This 2015 White Paper encompasses recommendations that emerged from the extensive discussions held during the workshop, and is aimed at providing the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to advance scientific excellence, improve quality and deliver better regulatory compliance. Due to its length, the 2015 edition of this comprehensive White Paper has been divided into three parts. Part 2 covers the recommendations for hybrid LBA/LCMS and regulatory agencies' inputs. Part 1 (small molecule bioanalysis using LCMS) and Part 3 (large molecule bioanalysis using LBA, biomarkers and immunogenicity) will be published in volume 7 of Bioanalysis, issues 22 and 24, respectively.

The Decennial Index of The White Papers in Bioanalysis: ‘A Decade of Recommendations (2007–2016)’

“…Prior to any cross-validation of LBA and LCMS techniques, it was recommended in 2014 to confirm that both technologies measure the same analyte [12]. It was not generally recommended to measure an analyte with both approaches; however, this may happen when one assay or technology runs into issues such as matrix interference, when early data are generated on one platform and then the platform is switched to the other; or when the molecular complexity necessitates additional characterization.…”

“…When cross-validating between LCMS and LBA, QC samples and incurred samples should be used, and the general acceptance criteria that should be applied are twothirds of sample results within 30% [9]. The following year, this recommendation was expanded to conclude that if cross-validation of LBA and LCMS data are required, then it is recommended to use the same capture reagent [12].…”

“…Potential binding molecules should be evaluated as part of an assay performance QC if it is judged that the assay could be impacted by the presence of these molecules. Capture with target, receptors and other reagents was proposed [12]. It was also recommended that the choices of instruments (HRMS or triple quadrupole), internal standards (SIL intact protein or SIL flanked peptide), extraction techniques and signature peptide should all be thoroughly evaluated to optimize method performance.…”

“…Discussing hybrid LBA/LCMS method development in 2015 concluded that existing LBA workflows can be used. Commercial LBA reagents can also be used [12]. Binding interferences can often be removed by using an acid pretreatment step.…”

“…These recommendations support original discussions in 2015, where it had been concluded that LCMS is best used as an orthogonal tool to assist when needed for ADA determination, for example, to overcome the lack of drug tolerance or other analytical interferences. LCMS can be used for immunoglobulin allotyping/isotyping by specific immunocapture of IgG, IgM and other immunoglobulin subclasses and/or by analysis of specific signature tryptic peptides reflecting immunoglobin subclasses [12].…”